RT-qPCR Extraction Control Red

A common practice in qPCR is to add a known amount of spiked control RNA after RNA extraction, this monitors PCR inhibition but has no value as an extraction control. The ideal situation is to have the test sample and internal control undergo the same processing prior to qPCR. Meridian has developed a RT-qPCR Extraction Control, which more closely mimics the test sample, as compared to spike controls. Genetic material from the test sample and the RT-qPCR Extraction Control is simultaneously extracted by common extraction methods, with the extraction control being as sensitive to inhibition and extraction failure as the test sample. Artificial RT-qPCR Extraction Control cells are of a known concentration, containing the Internal Control RNA sequence. This sequence contains no known homology to any organism and, importantly, has minimal interference with detection of sample RNA. The RT-qPCR Extraction Control cells are spiked into the lysis buffer with the target sample, prior to RNA extraction. Control Mix, which includes primers and probe, is then added to the reaction mix before amplification. Signal derived from the Internal Control RNA confirms the success of the extraction step. RT-qPCR Extraction Control also monitors co-purification of PCR inhibitors that may cause biased or false amplification patterns.